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Preparation of tissue sections using fluorescence-based detection

a tissue and fluorescence-based detection technology, applied in the field of tissue preparation, can solve the problems of inaccurate and inconsistent data provided by methods, and it is difficult to distinguish tissue from paraffin in an ffpe sample using existing methods

Active Publication Date: 2019-12-05
AGILENT TECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present patent provides a method for determining the location of a tissue in an embedded sample using a process called fluorescence emission. This method involves irradiating the embedded sample with light of a specific wavelength and detecting the emitted fluorescence. The location of the tissue can then be determined based on this fluorescence emission. The patent also provides an apparatus for slicing a tissue section from the embedded sample using a microtome and a light source. Overall, this technology allows for more precise and accurate analysis of embedded samples and improved tissue sectioning.

Problems solved by technology

Many existing methods provide inaccurate and inconsistent data when used to analyze different tissue and paraffin types, since many methods are sensitive to variability of optical and surface characteristics of tissue and paraffin.
In some cases, it is quite difficult to distinguish tissue from paraffin in an FFPE sample using existing methods.

Method used

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  • Preparation of tissue sections using fluorescence-based detection
  • Preparation of tissue sections using fluorescence-based detection
  • Preparation of tissue sections using fluorescence-based detection

Examples

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example 1

[0083]This Example illustrates a method of measuring fluorescence of endogenous fluorophores in human tissue in accordance with an embodiment of the present disclosure.

[0084]An FFPE human tissue block was obtained. The FFPE block was analyzed to determine if autofluorescence can be used to differentiate tissue in the FFPE block from paraffin. The uncut FFPE block was subjected to light corresponding to the excitation wavelengths of elastin and collagen. Any resulting fluorescence emission was detected using digital imaging. The FFPE block was cut using a ThermoFisher microtome blade, where consecutive cuts of the FFPE block were performed in intervals ranging from 5 to 40 microns in thickness. After each cut, the block face of the FFPE block was subjected to light corresponding to the excitation wavelengths of elastin and collagen and the fluorescence emission was detected using digital imaging. The images were collected at a 4× magnification. The pixel intensity for each image was ...

example 2

[0088]This Example illustrates a method of contrasting tissue using fluorescence emission of paraffin in accordance with an embodiment of the disclosure.

[0089]A FFPE human tissue block was obtained. The FFPE block was analyzed to determine if fluorescence of paraffin can be used to differentiate tissue in the FFPE block from paraffin. In particular, the experiment was conducted to determine if an image could be obtained of paraffin fluorescence in the presence of the endogenous fluorophores in tissue. Flourescence intensity of tissue in a FFPE sample was measured using a 365 nm LED excitation source with an emission filter centered at 560 nm (55 nm bandpass). Flourescence intensity of the paraffin surrounding the tissue in an FFPE sample was measured with a 280 nm LED excitation source and an emission filter centered at 405 nm (20 nm wide bandpass).

[0090]To collect this data, both the excitation sources front illuminated the block transversely at oblique angles of between 10 and 20 ...

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Abstract

The present disclosure is directed to an improved method for distinguishing tissue from an embedding medium, such as paraffin in a formalin-fixed paraffin-embedded sample. The method involves the use of fluorescence of naturally-occurring species in tissue to determine the location of the tissue in the embedded sample. An embedded sample is generally excited by light of a selected wavelength, and the fluorescence emission at an emitted wavelength is used to locate the boundary or location of the tissue in the embedded sample.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application claims the benefit of U.S. Provisional Application No. 62 / 537,848, filed Jul. 27, 2017, the contents of which are incorporated by reference in their entirety.FIELD OF THE INVENTION[0002]The present disclosure relates to methods of preparing tissue sections from an embedded sample such as a formalin-fixed paraffin-embedded sample using fluorescence-based detection.BACKGROUND[0003]The formation of a formalin-fixed paraffin-embedded (FFPE) tissue block serves to preserve the morphology and cellular content of a tissue sample. Tissue processing generally involves placing an isolated tissue in formalin for a time period such as a few days, and then embedding the tissue in a paraffin wax. FFPE samples can be conveniently stored at room temperature for extended periods of time, and are especially useful for immunohistochemical staining and morphology analyses. FFPE samples may also be used for profiling gene expression and study...

Claims

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Application Information

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IPC IPC(8): G01N1/30G01N1/28G01N1/06G01N35/00G06T7/00
CPCG01N2001/068G01N1/286G01N1/06G06T2207/10152G06T2207/30024G01N35/00594G01N1/30G06T2207/10064G01N2001/2873G06T7/0012G01N21/6458G06T7/11G06T7/136
Inventor SCHLEIFER, KYLEBERNICK, KRISTIN BRIANAMCCAMPBELL, ADRIENNESAMPAS, NICHOLAS M.LIM, VICTOR
Owner AGILENT TECH INC